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Mammalian (cytosine-5) methyltransferases cause genomic DNA methylation and lethality in Drosophila

Nature Genetics volume 23, pages 363–366 (1999)Cite this article

Abstract

CpG methylation is essential for mouse development1 as well as gene regulation and genome stability2,3,4,5. Many features of mammalian DNA methylation are consistent with the action of a de novo methyltransferase that establishes methylation patterns during early development and the post-replicative maintenance of these patterns by a maintenance methyltransferase6. The mouse methyltransferase Dnmt1 (encoded by Dnmt) shows a preference for hemimethylated substrates in vitro , making the enzyme a candidate for a maintenance methyltransferase7,8. Dnmt1 also has de novo methylation activity in vitro 9, but the significance of this finding is unclear, because mouse embryonic stem (ES) cells contain a de novo methylating activity unrelated to Dnmt1 (ref. 10). Recently, the Dnmt3 family of methyltransferases has been identified and shown in vitro to catalyse de novo methylation11. To analyse the function of these enzymes, we expressed Dnmt and Dnmt3a in transgenic Drosophila melanogaster. The absence of endogenous methylation in Drosophila12,13 facilitates detection of experimentally induced methylation changes. In this system, Dnmt3a functioned as a de novo methyltransferase, whereas Dnmt1 had no detectable de novo methylation activity. When co-expressed, Dnmt1 and Dnmt3a cooperated to establish and maintain methylation patterns. Genomic DNA methylation impaired the viability of transgenic flies, suggesting that cytosine methylation has functional consequences for Drosophila development.

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Figure 1: Expression of mouse methyltransferases in Drosophila.
Figure 2: Expression of Dnmt3a, but not Dnmt, causes developmental defects in Drosophila.
Figure 3: Expression of Dnmt3a, but not Dnmt, causes CpG methylation of genomic DNA in Drosophila.
Figure 4: Dnmt functions as an enhancer of the Dnmt3a phenotype.

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Acknowledgements

We thank D. Biniszkiewicz for Dnmt cDNA; D. Schneider and M. Voas for help with pictures; I. Rebay for fly strains; and H. LeBlanc for helpful discussions. This work was supported by NIH grants GM39341 (to T.O.-W.) and 5-R35-CA44339 (to R.J). F.L. received a research fellowship from the Deutsche Forschungsgemeinschaft.

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Authors and Affiliations

  1. Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA

    Frank Lyko, Helena Kashevsky, Matthew Tudor, Mary-Ann Mastrangelo, Terry L. Orr-Weaver & Rudolf Jaenisch

  2. Department of Haematology, University of Wales College of Medicine, Heath Park, Cardiff, UK

    Bernard H. Ramsahoye

  3. Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Terry L. Orr-Weaver & Rudolf Jaenisch

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Correspondence to Rudolf Jaenisch.

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Lyko, F., Ramsahoye, B., Kashevsky, H. et al. Mammalian (cytosine-5) methyltransferases cause genomic DNA methylation and lethality in Drosophila. Nat Genet 23, 363–366 (1999). https://doi.org/10.1038/15551

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